不同宿主肝片吸虫DNA限制性片段长度多态性分析

Restriction Fragment Length Polymorphism in Fasciola hepatica Isolated from Different Host

本文首次对分离自黄牛、水牛和牦牛的肝片吸虫的基因组DNA，以BamHⅠ、BglⅡ、EcoRⅠ、HindⅢ和PStⅠ等6种限制性内切酶消化后进行琼脂糖电泳，分析比较它们基因组DNA的限制性片段长度多态性（RestrictionFragmentLengthPolymorphism，RFLP）；同时用地高车标记的牦牛肝片吸虫基因组DNA为探针对不同宿主肝片吸虫DNA进行Southern印迹杂交，限制性内切酸酶切结果显示只有两种限制性内切酶（BamHⅠ和EcoRⅠ）在寄生于牦牛的肝片吸虫中检测到多态性，而水牛肝片吸虫与黄牛肝片吸虫基因组DNA之间没有检测到多态性，根据此结果计算了牦牛肝片吸虫与黄牛肝片吸虫和水牛肝片吸虫基因型间的遗传距离，Southern杂交结果显示牦牛肝片吸虫基因DNA的杂交谱带与黄牛肝片吸虫、水牛肝片吸虫基因组DNA的杂交港带存在一定的差异，本文的实验结果表明牦牛肝片吸虫与黄牛肝片吸虫、水牛肝片吸虫的亲缘关系相对较远，而水牛肝片吸虫与黄牛肝片吸虫的亲缘关系较近。

Six restliction endonucleases,BamH Ⅰ,Bgl Ⅱ,EcoR Ⅰ,Hind Ⅲ,,Hpa Ⅰ and Pst Ⅰ were used to study the ge-nomic DNA restriction fragment length polymorphism (RFLP ) in Fasciola hepatica isolated from yak,buffallo and cattle,meanwhile genomic DNA digested with BamHI and EcoRI were hybridized with a genomic DNA,which was prepared from yak F.hepatica and labeled with ixgoxin,in Southern.blotting.Only two (BamHI and EcoRI)out of six restriction enzymes disclosed polymorphism within yak F.hepatica.The restriction patterns generated by other 4 of the enzymes in yakF.hepatica were identical with those in buffolla and cattle F.hepatica.The Southern blotting hybridization showed that buffalo F.hepatica and cattle F.hepatica genomic DNA had identical bands,yak F.hepatica had different bands.Results showed that genomic DNA of yak F.hepatica was somewhat different from genomic DNA of buffallo and cattle F.hepatica.